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目的探讨人参皂苷代谢产物Compound K(CK)对炎症因子TNF-α诱导的人支气管上皮细胞BEAS-2B分泌调节活化正常T细胞表达和分泌趋化因子(RANTES)的影响及其机制。方法 ELISA法测定CK对BEAS-2B细胞上清液中RANTES含量的影响;采用RT-PCR和蛋白质免疫印迹技术检测RANTES mRNA及蛋白的表达情况;采用荧光素酶报告基因系统检测CK对活化蛋白转录因子1(activator protein 1,AP-1)和糖皮质激素受体(glucocorticoid receptor,GR)转录抑制或转录激活的影响;运用GR拮抗剂米非司酮,验证CK对RANTES的抑制效应是否由GR介导。结果 3~30μmol/L的CK抑制了TNF-α诱导的RANTES分泌、mRNA及蛋白水平;CK抑制AP-1的转录激活,在BEAS-2B细胞中激活GR信号通路;并且,CK通过GR抑制了BEAS-2B细胞分泌RANTES。结论 CK能够抑制炎症因子TNF-α诱导的支气管上皮细胞分泌RANTES,其机制可能与激活GR、抑制AP-1对下游靶基因的调控有关。
Objective To investigate the effect and mechanism of Compound K (CK), a metabolite of ginsenosides, on the expression and secretion of chemokines (RANTES) regulated by inflammatory cytokine TNF-α induced by BEAS-2B in human bronchial epithelial cells. Methods The effect of CK on the content of RANTES in BEAS-2B supernatant was measured by ELISA. The expression of RANTES mRNA and protein was detected by RT-PCR and Western blotting. The luciferase reporter gene system was used to detect the transcription of activated protein 1, activator protein 1 (AP-1) and glucocorticoid receptor (GR). Using GR antagonist mifepristone, the inhibitory effect of CK on RANTES was tested by GR mediate. Results CK at 3 ~ 30μmol / L inhibited TNF-α-induced RANTES secretion, mRNA and protein levels; CK inhibited the transcriptional activation of AP-1 and activated GR signaling in BEAS-2B cells; BEAS-2B cells secrete RANTES. Conclusions CK inhibits the secretion of RANTES by bronchial epithelial cells induced by inflammatory cytokine TNF-α, which may be related to the activation of GR and the inhibition of AP-1 on the downstream target genes.